Search Results (2,123)

Human pluripotent stem cells (hPSCs) are a promising source for regenerative medicine due to their self-renewal and differentiation capacities. However, genetic instability acquired during reprogramming and in vitro culture presents major safety challenges for clinical translation. Recurrent mutations, especially structural variants (SVs), are of particular concern as they can impair differentiation and increase tumorigenic risk. In this review, we establish and systematically explore a central causal axis: SVs–three dimensional (3D) genome disruption–safety of hPSC-based therapy. We propose that SVs critically compromise therapeutic safety by perturbing the 3D architecture of the genome, leading to pathogenic rewiring of enhancer–promoter interactions. This rewiring, exemplified by “enhancer hijacking” and “enhancer loss,” can aberrantly activate oncogenes or silence tumor suppressors even in the absence of copy number variations. Thus, 3D genome disruption provides a key mechanistic explanation for SV-driven tumorigenic potential and impaired differentiation fidelity in hPSCs. By highlighting this causal axis, our review not only advances the mechanistic understanding of SV-associated risks but also provides actionable insights for the development of more rigorous quality standards for hPSC-based cell therapy products. Full article

Excitotoxicity is one of the factors that participates in neurodegeneration, impairing neuronal and glial cells’ function, and leading to the development of chronic neurodegenerative diseases. The main mechanism of action lies in the overstimulation of excitatory receptors, especially the NMDA (N-methyl-D-aspartic acid) receptor, by glutamate, which promotes a massive influx of Ca²⁺ that is not sufficiently buffered by the intracellular machinery, or not released by mechanisms such as Ca²⁺ ATPase and plasma membrane Ca²⁺/Na⁺ exchanger promoting, among other toxic effects, mitochondrial damage and an increase in reactive oxygen species (ROS). Notably, many cases reported of long COVID-19 describe significant brain alterations and neuropsychiatric disorders, including delirium, depression, etc., and patients required increased use of antidepressant or anxiolytic drugs, for example. In addition, emerging evidence links neurodegeneration as a potential long-term sequelae associated with an increased number of patients with cognitive disorders. This review analyzes data from the literature regarding brain alterations associated with post-COVID-19 syndrome and explores a potential link to the excitotoxicity pathways, due to its participation in neurodegeneration by homeostatic failure, and it is clearly present in various brain conditions, such as Alzheimer’s and Parkinson’s diseases. Full article

Goose blood has anticancer properties and was recorded in ancient China, but the specific molecular mechanisms underlying this effect still require further exploration. In this study, SW1990 cells were treated with goose serum or plasma, and transcriptome analysis was performed to explore the function of goose blood on cancer cells. Metabolomic profiling was also performed on goose serum, goose plasma, chicken serum, and chicken plasma to identify the bioactive substances responsible for the anticancer effect. The study examined the effects of goose plasma and serum on SW1990 cells and compared the metabolites between goose and chicken blood. Wound scratch, CCK-8, and Annexin V-PI assays showed that goose plasma and serum inhibited SW1990 cell proliferation at 24 and 48 h. Both treatments reduced cell viability, with serum inducing early and late apoptosis and plasma inducing late apoptosis. RNA sequencing (RNA-seq) identified 2259 (1418 upregulated, 841 downregulated) and 2731 (1844 upregulated, 887 downregulated) differentially expressed genes (DEGs) in the plasma and serum groups versus the negative control (NC), respectively, and 689 DEGs between the plasma and serum groups. Gene Ontology (GO) and KEGG pathway analyses revealed that the DEGs were enriched in processes such as lipid metabolism, JAK-STAT, and IL-17 pathways. Untargeted liquid chromatography–tandem mass spectrometry (LC-MS/MS) analysis identified distinct metabolites in goose and chicken blood, with unique metabolites and differential ones between groups. In SW1990 cells, four metabolite subclusters matched the plasma and serum effects. In summary, goose blood can suppress cancer cells by regulating gene expression to affect the key signaling pathways involved in cancer cell apoptosis and autophagy. Certain metabolites present at high concentrations in goose blood, such as cucurbitacin D and Oleoyl-L-carnitine, may also contribute to the inhibition of cancer cell proliferation and migration. These findings suggest that goose blood holds broad application prospects as a future auxiliary drug for cancer treatment, and this study provides a theoretical basis for the further application of goose products. Full article

Cadmium (Cd) is a persistent environmental pollutant that poses a significant health risk to humans and animals, with acute exposure known to induce kidney injury. Fucoidan (Fc), a natural bioactive polysaccharide derived from brown algae, exhibits diverse biological activities; however, its potential to protect against Cd-induced kidney damage and the underlying mechanisms remain unclear. In this study, we investigated the effects of Fc on Cd-induced renal injury in vitro and further explored the role of transcription factor EB (TFEB) in regulating autophagy in its protective mechanism. Our results demonstrate that in Cd-exposed porcine kidney cells (PK-15), Fc suppressed the expression of renal inflammatory factors (TNF-α, IL-1β) and kidney injury markers (NGAL, NTN-1, KIM-1), reduced reactive oxygen species (ROS) production, and downregulated apoptosis-related proteins (cleaved caspase-3 and cleaved caspase-9). Mechanistically, Fc upregulated TFEB protein expression, enhanced the levels of lysosomal function-related proteins (Cathepsin B, CTSB; Cathepsin D, CTSD), and reversed Cd-induced autophagic flux blockade. Importantly, TFEB silencing abolished the protective effects of Fc. Collectively, these findings suggest that Fc exerts renoprotective effects against Cd-induced injury by restoring autophagic flux, a process that involves TFEB. Full article

Spi-1 Proto-Oncogene (SPI1) encodes Purine-rich box 1 Transcription Factor (PU.1), a transcription factor of the ETS family that regulates hematopoietic lineage commitment and immune cell differentiation. Alteration of PU.1 dose or ETS domain integrity may interfere with transcriptional programs, which adds to hematopoietic dysregulation and leukemogenesis. Even though changes in SPI1 expression have been associated with acute myeloid leukemia (AML), the structural and regulatory effects of missense mutations at the PU.1 ETS domain have not been entirely studied, and targeting the PU.1 ETS domain by ligands is an area of computational analysis that should be further pursued. To computationally describe deleterious missense variants of SPI1 in terms of structural stability, evolutionary conservation, post-translational modification (PTM) context and interaction networks, and to measure ADMET-mediated molecular docking of cinnamic acid with the PU.1 ETS domain (8EQG) as a potential modulator. Missense nsSNPs were obtained through Ensembl and narrowed down by consensus prediction of pathogenicity (PredictSNP, combining SIFT, PolyPhen, SNAP and PhD-SNP and other tools). InterPro/UniProt was used for domain mapping. SWISS-MODEL was used to produce wild-type and mutant PU.1 versions, which were analyzed on the structural alignment and Cα–Cα displacement parameters in UCSF Chimera (v1.19). The estimation of stability change was carried out with I-Mutant and MUpro. Prediction of PTM sites was done using MusiteDeep and exploration of functional partners was done using STRING. Human, mouse and zebrafish orthologue conservation was measured by means of MAFFT alignment. GEPIA2 was used to compare the expression of SPI1 in AML (TCGA-LAML) and normal tissues (GTEx). AutoDock Vina (grid center 6, −2, −9 A; 20 × 20 × 20 A; 16 exhaustiveness) was used to prepare cinnamic acid and dock it into the PU.1 ETS domain (8EQG), with SwissDock being used for consistency checks. SwissADME and ADMETlab 2.0 were used to predict drug-likeness, pharmacokinetics, and toxicity. Nine missense mutations were routinely considered as deleterious with the majority of them being located in or near the ETS DNA-binding domain. Structural comparisons showed local perturbations of the structure and I189F and H211P yielded the greatest conformational changes between prioritized variants whereas other forms had minimal movements. A predominantly destabilizing trend was supported by stability prediction whereby V241G had the strongest destabilization signal with further destabilizations being predicted in I189F and R259C. PTM mapping revealed several potential regulatory residues (phosphorylation, acetylation, ubiquitination, and methylation), which indicated that there could be crosstalk between the sequence variation and the transcriptional regulation. The SPI1 was placed in a central hematopoietic transcriptional module (containing RUNX1, CEBP members, GATA1 and IRF factors) by the STRING network. The cross-species alignment showed that there was high conservation of a number of the mutation sites, which would support functional constraint at the ETS region. The expression analysis revealed that the level of SPI1 mRNA in AML was significantly elevated compared to normal tissues. Docking also indicated a slight and reproducible interaction of cinnamic acid with the ETS domain (top affinity −4.27 kcal/mol), with a solitary leading polar anchor and supportive hydrophobic interactions, which is akin to interaction between fragments. The ADMET profiling revealed the likelihood of success in the oral drug-likeness and low CYP inhibition liability, as well as signifying the presence of a possible hepatotoxicity signal that needs further confirmation through experiments. Comprehensive computational studies suggest that certain pathogenic variants of SPI1 missense defects, especially in the ETS domain, can result in loss of PU.1 structural stability and regulatory environment, which are in line with the disturbed hematopoietic regulation and AML-related dysregulation. Cinnamic acid demonstrates moderate yet reproducible binding to the PU.1 ETS domain and has an overall favorable developability profile, which indicates that it is better considered as a starting scaffold, as opposed to an active inhibitor. The results give a logical basis of focused biochemical validation and structure-directed optimization of ETS domain modulators in hematologic disease settings. Full article

This study aimed to explore the alleviating effects of fisetin, a polyphenolic flavonoid, on ovarian dysfunction in a D-galactose (D-gal)-induced aging mouse model, as well as the underlying mechanisms, using both in vivo and in vitro experiments. Mice were subcutaneously injected with D-gal (100 mg/kg/day) for 60 days to establish the ovarian aging model; during the final 30 days, fisetin (10, 20, 30 mg/kg/day) was given orally. In addition, a senescent model of granulosa cell (GC) was established using D-gal and treated with fisetin. Fisetin supplementation improved ovarian endocrine function and reproductive capacity in aging mice, as reflected by regularized estrous cycles, elevated estradiol levels, and increased embryo numbers. Furthermore, fisetin reduced the number of atretic follicles and the extent of ovarian fibrosis and senescence, while simultaneously restoring the proliferation-apoptosis balance in follicular GCs, as well as alleviating oxidative stress. RNA-sequencing revealed that AMP-activated protein kinase (AMPK)/mechanistic target of rapamycin (mTOR) signaling and mitophagy were involved in the protective effects of fisetin against ovarian aging. Consistently, fisetin treatment promoted mitophagy, accompanied by AMPK/mTOR activation in ovarian tissues and GCs following D-gal exposure. Inhibition of AMPK attenuated the effect of fisetin on mitophagy. Additionally, blockage of mitophagy also reversed the beneficial effects of fisetin on mitochondrial injury, oxidative stress, cell cycle arrest, and cellular senescence in D-gal-induced senescent GCs. These findings indicate that fisetin prevents ovarian aging by suppressing follicular GC oxidative damage and ameliorating cell cycle arrest via activation of AMPK/mTOR-mediated mitophagy, thereby preserving female fertility. Full article

Deep learning has demonstrated high efficiency in histopathological image analysis, particularly in lung cancer classification. However, the stability of these models with image corruption and cross-dataset validation remains an important practical concern. In this study, we explored the potential of adding spectral information derived from the discrete wavelet transform (DWT) and spatial convolutional representations to enhance the robustness of multi-class lung cancer classification between Normal, Adenocarcinoma and Squamous cell carcinoma. The lightweight ResNet18 backbone was used to obtain spatial features, and spectral descriptors were obtained through wavelet sub-bands and integrated through early feature-level fusion. The models were trained and evaluated using the LC25000 dataset. Subsequently, it was tested under controlled perturbations, such as Gaussian noise and Gaussian blur. Three random seeds were used to assess performance variability, and paired t-tests were conducted as an indicative statistical measure of the results. Under clean conditions, the spatial and hybrid models were nearly saturated, and there was no significant difference between them (spatial: 99.85 ± 0.26; hybrid: 99.72 ± 0.22; p = 0.1217). The hybrid model exhibited higher robustness when Gaussian noise (σ = 0.05) was added, which resulted in 84.89% ± 4.52% accuracy versus 74.99% ± 7.20% of the spatial baseline (p = 0.0443) with an observed effect size (Cohen’s d = 2.64), noting that these estimates are based on a limited number of runs and should be interpreted with caution. The same behavior was observed in Gaussian blur perturbations, where the hybrid representation was slightly more stable. We also investigated a simplified adaptive gating mechanism process and found that the learned gate parameter also tends to converge towards spatial feature dominance with a model trained with clean data. Finally, cross-dataset validation with LungHist700 showed a slight increase in the balanced accuracy of the hybrid model (0.5158) over the spatial baseline (0.4722). These results indicate that spectral and spatial features can be used to enhance robustness to image corruption and still yield high classification accuracy, indicating that spectral–spatial representations can improve robustness under controlled perturbations, whereas their impact on cross-dataset generalization remains limited. The results further indicate that robustness improvements are strongly influenced by training strategies, such as noise augmentation, whereas the contribution of fusion is comparatively moderate. Full article

The tissue regeneration of sea cucumber (Apostichopus japonicus) involves precise intercellular signal pathway transduction and gene expression regulation. This study investigated the function of the Notch signaling pathway in A. japonicus papilla regeneration and its modulation by LED light of varying intensities. We detected the expression patterns of Notch signaling pathway-related genes and their downstream cell proliferation-related genes during papilla regeneration, and further verified the pathway function via gene silencing, combined with histological analyses to explore LED-mediated effects. Gene expression assays revealed that AjNotch, AjSu(H), AjHes1, AjCyclinA, AjCyclinD and AjCDK8 were significantly upregulated at 28 days post papilla excision (p < 0.05). LED light treatment accelerated papilla regeneration in a light intensity-dependent manner, with the most pronounced promotion at 2000 lx (p < 0.05). Moreover, LED light treatment was associated with altered expression of Notch signaling pathway genes and their downstream proliferation-related genes in a light intensity-dependent manner. Gene silencing of AjNotch significantly downregulated its downstream target genes (p < 0.05), attenuated the regenerative promotion of LED light, and reduced cell proliferation rate (p < 0.05). These findings suggest that the Notch signaling pathway is pivotal for A. japonicus papilla regeneration, and LED light modulates papilla regeneration with concurrent changes in the expression of Notch pathway-related genes. This study provides novel insights into the function of the Notch signaling pathway in echinoderm regenerative development. Full article

Although sorafenib (SOR) is effective for advanced hepatocellular carcinoma (HCC), significant metabolic heterogeneity limits its therapeutic effect. In this study, we employed high-resolution matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) to profile the spatial lipidomic alterations in 3D HepG2 spheroids following SOR treatment. Interestingly, sphingophospholipid and glycerophospholipid metabolism played crucial roles. In an orthotopic HCC mouse model, immunohistochemical and immunofluorescence staining confirmed that SOR induced immunological and inflammatory changes. Moreover, transcriptomic and Q-PCR analyses showed increased expression of Stat1, Zbp1, Parp14, Irf1, and Tifa along with decreased Eif4e2 in the SOR treatment group compared to the tumor control group. Bio-layer interferometry and molecular docking data also indicated that ZBP1 possessed favorable binding affinities with SOR. Overall, our findings demonstrated that SOR dramatically disrupted sphingolipid metabolism in tumor cell spheroids and, in an orthotopic model, activated the NOD-like receptor signaling pathway, accompanied by altered secretion of inflammatory factors and macrophage polarization. These results suggest that SOR exerts dual effects on tumor cell lipid metabolism and the tumor immune microenvironment. These findings provide a conceptual basis for future exploration of lipid-modulating therapeutic strategies in HCC. Full article

Diamond-like carbon (DLC) coatings are increasingly explored as a practical route to enhance the surface performance of biomedical implants and tissue engineering scaffolds, particularly when combined with additive manufacturing. Rather than serving only as protective layers, DLC coatings allow for independent tuning of surface properties without modifying the bulk structure, which is especially relevant for complex 3D-printed components. This flexibility is often what makes them attractive for biomedical design. This review is structured around two main application areas: DLC coatings for prosthetic implants and DLC coatings for tissue engineering scaffolds. Within this context, the influence of DLC structure (e.g., sp²/sp³ bonding, hydrogen content, and doping) on mechanical, tribological, and biological behavior is discussed. Particular attention is given to additively manufactured metallic implants and porous scaffolds, where large surface area and internal architectures complicate coating uniformity and adhesion. Reports show that DLC coatings can improve corrosion resistance, reduce wear, and influence biological responses, such as antibacterial activity and cell interactions. Several challenges remain to be solved, especially in achieving uniform coating penetration in porous networks and in ensuring long-term stability under physiological conditions. The combination of additive manufacturing and DLC coatings has been shown to offer the potential to become an enabling technology for next-generation biomedical devices. Full article

Melia azedarach L. is a plant known for its traditional medicinal uses. Limonoids (triterpenes), which have a wide range of pharmacological effects, are the most critical active ingredients; however, their potential effects on liver cancer remain to be further explored. In this study, seven limonoids were isolated from the bark of Melia azedarach, including two new compounds, 11α-hydroxy-12-Oxo-Meliarachin I (1) and 29-Oxo-12-dehydroneoazedarachin D (3), along with five known compounds (2, 4–7), to evaluate their effect on liver cancer in vitro. The results showed that compounds 1–7 exhibited varying degrees of inhibitory effects on Hep3B cells. Among these, compound 6, Isotoosendanin (ITSN), displayed the most potent activity, with an IC₅₀ value of 15.06 μg/mL. Mechanism studies have shown that ITSN inhibits cell proliferation and promotes apoptosis in Hep3B cells. It induces reactive oxygen species (ROS) accumulation to trigger oxidative stress injury, suppresses the activation of the MAPK and PI3K/AKT signaling pathways, further activates the p53 pathway to induce cell cycle arrest, and ultimately initiates the apoptotic cascade. ITSN can also inhibit tumor growth in immunodeficient mice receiving allogeneic transplantation. In summary, we systematically studied the limonoids in the bark of Melia azedarach and elucidated the anti-hepatocarcinoma activity of ITSN in vitro and in vivo, providing promising evidence for its potential use as a natural active ingredient in the prevention and treatment of cancer. Full article

Background: Microfibrillar-associated protein 2 (MFAP2) is implicated in various malignancies, yet its specific role and molecular mechanisms in glioblastoma (GBM) progression remain poorly understood. Methods: We analyzed MFAP2 expression in human clinical specimens and murine models. Functional impacts were assessed in U251 cells via gain- and loss-of-function assays. Mechanistic studies explored the interplay between autophagic flux and Wnt/β-catenin signaling. An orthotopic GL261 syngeneic orthotopic model validated these findings in vivo. Results: MFAP2 was significantly overexpressed in GBM, correlating with poor patient prognosis. In vitro, MFAP2 markedly enhanced U251 viability, migration, and invasion while suppressing apoptosis. Mechanistically, MFAP2 triggered autophagic flux, subsequently activating the Wnt/β-catenin cascade and its downstream targets (MMP9, c-Myc, Cyclin D1). Pharmacological inhibition of either autophagy or Wnt signaling effectively abrogated these oncogenic phenotypes. In vivo, MFAP2 knockdown reduced tumor volume by 62.4% and suppressed the autophagy–Wnt axis. Conclusions: MFAP2 is an oncogenic regulator in glioblastoma models that links autophagy activity to Wnt/β-catenin signaling. Our findings support MFAP2 as a candidate prognostic biomarker and a potential therapeutic target; however, additional validation in larger molecularly annotated clinical cohorts and multiple GBM models is warranted. Full article

Epidemiological studies have demonstrated that kidney aging is a risk factor for acute kidney injury (AKI) and chronic kidney disease (CKD). Therefore, understanding the mechanisms of kidney aging is key to designing novel anti-kidney aging strategies. In this regard, animal models of kidney aging are essential tools. In this review article, we focus on D-galactose (D-gal)-induced accelerated aging in rodents. This animal aging model is a popular and widely used experimental method in the field of aging and aging-related degenerative disorders. It has been shown that the major characteristics of the D-gal-induced aging process are increased oxidative stress, decreased antioxidant enzymes, elevated cell death, increased tissue fibrosis, and accumulation of inflammatory mediators. This review focuses on D-gal-induced kidney aging in mice and rats, with discussions on both kidney aging mechanisms and anti-kidney aging regimens using this model. It is our belief that D-gal induction of accelerated kidney aging will continue to be used as a convenient platform for elucidating kidney aging mechanisms and exploring novel anti-kidney aging targets that may slow down kidney aging and retard the development of aging-related renal disorders. Full article

Cancer exhibits pronounced heterogeneity at both spatial and cellular levels, contributing to variability in therapeutic responses and the emergence of treatment resistance. This heterogeneity is underscored by the diverse genetic, epigenetic, and phenotypic variations found within tumor cell populations. Cancer stem cells (CSCs), although representing a minor fraction of tumor cells, possess the capacity to self-renew and differentiate, thereby driving the dynamic evolution of tumor heterogeneity. CSCs interact intricately with various elements of the tumor microenvironment (TME), further amplifying this heterogeneity. Recent advancements in organoid technology have facilitated the development of CSC-derived organoid models that more faithfully recapitulate the TME and intratumoral heterogeneity, which conventional 2D culture systems fail to replicate. These CSC-derived organoid systems not only preserve the structural and genomic characteristics of tumors, but they also enable the exploration and evaluation of therapeutic strategies that reflect tumor complexity. However, CSC-derived organoid systems face several challenges, such as the rarity of CSCs, lack of standardized culture conditions, absence of TME components, limited predictive accuracy, and insufficient modeling of tumor heterogeneity. This review discusses these limitations and explores potential solutions, including the use of artificial intelligence (AI) to enhance treatment predictability. These innovations may improve the utility of organoid models for therapeutic evaluation and for targeting tumor heterogeneity. Ultimately, CSC-derived organoids may serve as a valuable platform for advancing precision medicine and cancer research. Full article

Background: Esophageal squamous cell carcinoma (ESCC) remains a highly lethal malignancy with limited therapeutic options, in part due to frequent activation of nuclear factor erythroid 2-related factor 2 (NFE2L2 or NRF2). Gain-of-function mutations in NRF2 disrupt its negative regulation by Kelch-like ECH-associated protein 1 (KEAP1), resulting in sustained NRF2 signaling that promotes tumor growth and resistance to chemotherapy and radiation. We previously identified the FDA-approved drug pyrimethamine (PYR) as an NRF2 inhibitor and demonstrated that inhibition of dihydrofolate reductase (DHFR) represents the primary mechanism underlying its NRF2-suppressive activity, supporting its advancement into a Phase I window-of-opportunity clinical trial (NCT 05678348). Meanwhile, in NRF2^W24C-KYSE70 and NRF2^D77V-KYSE180 cells, PYR promoted NRF2^Mut ubiquitination and proteasomal degradation and shortened its half-life. This study aims to explore additional modes of action by which PYR inhibits NRF2. Methods: Cell cycle analysis was performed by flow cytometry. Cell proliferation, apoptosis and chemosensitivity were assessed by Live-Cell Analysis System, while radiosensitivity was evaluated using X-ray irradiation and the CellTiter-Glo assay. Molecular interactions between NRF2 and KEAP1 were examined through Co-IP and PLA, and the direct binding of PYR to KEAP1 was quantified using ITC and SPR. Molecular docking and dynamic simulations were employed to predict potential PYR-binding pockets within the Kelch domain. Results: Using genetically defined isogenic ESCC cell models, we show that activation of mutant NRF2 (NRF2^Mut) or wild-type NRF2 (NRF2^WT) produces distinct, context-dependent effects on squamous differentiation, proliferation, and therapeutic response. We further demonstrate that PYR restores sensitivity to chemotherapy and ionizing radiation in NRF2^Mut ESCC cells. Mechanistically, short-term PYR treatment promotes KEAP1-dependent proteasome-mediated degradation of NRF2^W24C. Biochemical and biophysical assays indicate that PYR enhances the interaction between KEAP1 and NRF2^W24C in a manner associated with KEAP1-dependent proteasomal degradation. Computational modeling further suggests that PYR may engage a pocket within the Kelch domain to facilitate the NRF2^W24C-KEAP1 interaction. Conclusions: These findings show that PYR functionally restores KEAP1-mediated NRF2 degradation of select NRF2^Mut through a glue-like effect and overcomes therapy resistance in ESCC. Although the proposed glue-like mechanism remains hypothetical, this work supports further investigation into the NRF2–KEAP1 interaction and may inform the development of KEAP1-targeted strategies for NRF2^Mut cancers, including ESCC. Full article